Invitrogen

ARF6 Polyclonal Antibody

Product Image Gallery

ARF6 Antibody — Fig 6 Arf6 is recruited by R . typhi RalF and is required for infection. (A) Ectopically expressed RalF RtFL co-localizes with Arf6 but not Arf5. HeLa cells co-expressing EYFP, EYFP-RalF RtFL or EYFP-RalF RbFL and mRFP-Arf6 (left) or -Arf5 (right) were fixed with 4% para-formaldehyde. Nuclei were stained with DAPI (blue). (Scale bar: 10 mum). (B) RalF RtFL pull-down of Arf6. Lysates from HEK293T cells expressing mRFP-Arf5 or -Arf6 were incubated with HisPur Cobalt resin bound with rHis-RalF RtFL or resin alone. Bound proteins were eluted with imidazole and analyzed by protein immunoblot using antibodies as indicated. (C) Arf6 is recruited during R . typhi entry. HeLa cells expressing mRFP-Arf5 or -Arf6 (red) were infected with partially purified R . typhi (MOI ~100). Ten minutes post infection, cells were fixed and R . typhi detected with anti- R . typhi serum (green). DAPI (blue) is shown in the merged image. Boxed regions are enlarged to show detail. White arrowheads indicate R . typhi . (Scale bar: 5 mum). (D) Arf6 knockdown inhibits R . typhi infection. HeLa cells transfected with negative, Arf6, or Arf5 siRNA were infected with partially purified R . typhi (MOI ~100). At 2 hrs post infection, cells were fixed, plasma membrane stained with Alexa Fluor 594 wheat germ agglutinin, and R . typhi detected with rat anti- R . typhi serum and Alexa Fluor 488 anti-rat antibody. The number of R . typhi per host cell was counted for 100 host cells for three independent experiments.

ARF6 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of Arf6 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 5% normal goat serum (Product # 31873) for 15 minutes at room temperature. Cells were probed with an Arf6 polyclonal antibody (Product # PA1-093) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

ARF6 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on normal biopsies of deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Rabbit Polyclonal Antibody recognizing Arf6 (Product # PA1-093) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

ARF6 Antibody in Western Blot (WB) — Western blot analysis of Arf6 was performed by loading 50 µg of various whole cell lysate onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were then probed with a rabbit polyclonal antibody recognizing Arf6 (Product # PA1-093) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were then washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody (Product # 31460) at a dilution of 1:10000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Super Signal West Pico (Product # 34080).

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

1:20-1:100

Immunofluorescence (IF)

1:20 - 1:100

Immunohistochemistry (Paraffin) (IHC (P))

1:20-100

Western Blot (WB)

1:1000

Published Applications

Miscellaneous PubMed (MISC)

See 1 publication below

Western Blot (WB)

See 2 publications below

Immunocytochemistry (ICC)

See 1 publication below

Immunofluorescence (IF)

See 1 publication below

Product Specifications

Species Reactivity

Human, Mouse, Non-human primate

Published species

Human

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

C-terminal human recombinant protein fragment

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

PBS with 1mg/mL BSA, 30% glycerol

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2539860

Target Information

ADP-ribosylation factor 6 (ARF6) is a guanine nucleotide-binding protein involved in protein trafficking and the regulation of endocytic recycling and cytoskeleton remodeling. As part of the RAS superfamily, Arf-6 stimulates the ADP-ribosyltransferase activity of cholera toxin and play a role in vesicular trafficking and as activators of phospholipase D. It may also be involved in dendritic spine development and the regulation of dendritic branching and filopodia extension.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.